Substance containing proteins and insoluble dietary fibers derived from the germinated seed of a grass family plant and uses thereof

ABSTRACT

There is provided a substance which was isolated from the germinated seed of a grass family plant and which contains proteins and insoluble dietary fibers. There are also provided a pharmaceutical composition comprising as an active ingredient said substance which was isolated from the germinated seed of a grass family plant and which contains proteins and insoluble dietary fibers, and a food composition comprising said substance which was isolated from the germinated seed of a grass family plant and which contains proteins and insoluble dietary fibers, as well as uses of said substance.

TECHNICAL FIELD

[0001] The present invention relates to substances which are isolatedfrom the germinated seed of a grass family plant and which containproteins and insoluble dietary fibers, and uses thereof

BACKGROUND ART

[0002] Ulcerative colitis has been rapidly growing in the number ofpatients in Japan since the 1970's. Main complaints of the diseaseinclude bloody diarrhea, abdominal pains and the like. It is a chronicand diffuse disease of the large intestine, and, after the initial onsetat the rectum, it expands in an ascending manner to the deep part of thelarge intestine with repeated remissions and flare-ups. The etiology ofthe disease is unknown at present although its association with the diethas been studied because the incidence of the disease increased with theWesternization of eating habits of the Japanese people. Some researcherssuggest that disease may be an autoimmune one, but no proposals on theetiology have obtained public acceptance so far. Very few drugs knowntoday are effective for the treatment of the disease except that asalazosulfapyridine is considered to have a therapeutic effect on mildcases of the disease. However, this drug is an antibiotic which isadministered in large quantities and thereby may cause problematic sideeffects such as diarrhea.

[0003] Even a possible therapeutic drug for ulcerative colitis currentlyunder development is a mere improvement of the salazosulfapyridine, andthe problem of its side effects has not been completely solved.

[0004] Incidentally, cancer ranks top as a cause of deaths in Japantoday. It is an extremely intractable disease and the ratio of itssuccessful treatment is not very high at present. Current treatments ofcancer include surgical removal, radiation exposure, chemotherapy(administration of anti-cancer agents), immunological therapy, etc.,with great strides being made in each method. However, cancer treatmentby a single method is believed to be a very difficult challenge.

[0005] The surgical process is a very effective means since it removesthe tumor cells themselves but its sole use cannot attain a completecure and, hence, radiation therapy and chemotherapy are simultaneouslyapplied in most cases. The biggest problem encountered in this case isthe side effects resulting from the radiation therapy and thechemotherapy. Either therapy attacks cancerous cells having highproliferating activities, so naturally severe damages are also inflictedupon tissues having high rate of growth such as mucosal epithelialcells. As a result, the intestinal mucosa is greatly damaged therebycausing a significant reduction in the absorption of orally administerednutrients and an ensuing severe diarrhea, which ultimately put thepatient in a greatly damaged condition. In an extreme case, the patientmay eventually die. However, very few effective means have so far beenavailable that can protect the damage of intestinal mucosa and preventdiarrhea.

[0006] If the bowels of patients suffering from colon cancer, Crohn'sdisease, etc., are extensively excised, post-operative dyspepsia andinsufficient absorption of electrolytes may cause the patients to havedifficulty in normal bowel movement resulting in an extremely highincidence of diarrhea etc., and this can badly affect the patients inleading their normal social life. The same problem is shared by patientswho were forced to have an artificial anus for some reason. As a meansto solve these problems, there may be mentioned an oral liquid diet thatleaves very little residue after absorption. However, the oral liquiddiet causes many problems on the part of patients, such as economicburdens, some pains encountered in uptake, and mental burdens due toregulated diets. Thus, there has been a great demand for an effectivemeans that can alleviate these burdens.

[0007] Thus, it is an object of the present invention to provide safematerials that can solve the problems stated above.

[0008] It is another object of the present invention to providepharmaceutical compositions and food compositions comprising saidmaterial.

[0009] It is a further object of the present invention to provide amethod of treating ulcerative colitis.

[0010] It is a still further object of the present invention to providea method of preventing or reducing the side effects of cancertreatments.

[0011] It is yet another object of the present invention to provide amethod of improving the bowel movement of patients who have undergoneintestinal ablation or patients with an artificial anus.

DISCLOSURE OF THE INVENTION

[0012] After the intensive research conducted to solve theabove-mentioned problems, the inventors have discovered that a substancewhich was separated from the germinated seed of a grass family plant andwhich contains proteins and insoluble dietary fibers (hereinafterreferred to as “the substance containing protein and insoluble dietaryfibers”) can attain the above objects and thereby have completed thepresent invention. The gist of the present invention is as follows:

[0013] (1) a substance which was isolated from the germinated seed of agrass family plant and which contains proteins and insoluble dietaryfibers;

[0014] (2) a pharmaceutical composition comprising as an activeingredient a substance which was isolated from the germinated seed of agrass family plant and which contains proteins and insoluble dietaryfibers;

[0015] (3) a food composition comprising a substance which was isolatedfrom the germinated seed of a grass family plant and which containsproteins and insoluble dietary fibers;

[0016] (4) a method of treating ulcerative colitis comprisingadministering to a patient with ulcerative colitis an effective amountof a substance which was isolated from the germinated seed of a grassfamily plant and which contains proteins and insoluble dietary fibers;

[0017] (5) a method of preventing or reducing the side effects of cancertreatments comprising administering to a patient with a cancer aneffective amount of a substance which was isolated from the germinatedseed of a grass family plant and which contains proteins and insolubledietary fibers;

[0018] (6) a method of improving the bowel movement of a patient who hasundergone intestinal ablation comprising allowing the patient to take aneffective amount of a substance which was isolated from the germinatedseed of a grass family plant and which contains proteins and insolubledietary fibers;

[0019] (7) a method of improving the bowel movement of a patient with anartificial anus comprising allowing the patient to take an effectiveamount of a substance which was isolated from the germinated seed of agrass family plant and which contains proteins and insoluble dietaryfibers;

[0020] (8) use as a pharmaceutical drug of a substance which wasisolated from the germinated seed of a grass family plant and whichcontains proteins and insoluble dietary fibers;

[0021] (9) use, for the treatment of ulcerative colitis, of a substancewhich was isolated from the germinated seed of a grass family plant andwhich contains proteins and insoluble dietary fibers;

[0022] (10) use, for preventing or reducing the side effects of cancertreatments, of a substance which was isolated from the germinated seedof a grass family plant and which contains proteins and insolubledietary fibers;

[0023] (11) use as food of a substance which was isolated from thegerminated seed of a grass family plant and which contains proteins andinsoluble dietary fibers;

[0024] (12) use, for improving the bowel movement of a patient who hasundergone intestinal ablation, of a substance which was isolated fromthe germinated seed of a grass family plant and which contains proteinsand insoluble dietary fibers; and

[0025] (13) use, for improving the bowel movement of a patient with anartificial anus, of a substance which was isolated from the germinatedseed of a grass family plant and which contains proteins and insolubledietary fibers.

BRIEF EXPLANATION OF THE DRAWINGS

[0026]FIG. 1 shows the diarrhea scores of Experiment Group 1 andComparative Group 1.

[0027]FIG. 2 is a pair of photographs showing the biological morphologyof the appearance of stools from Comparative Group 1 (2-1) andExperiment Group 1 (2-1).

[0028]FIG. 3 is a pair of photographs showing the biological morphologyof the appearance of the colon mucosas of Comparative Group 1 andExperiment Group 1.

[0029]FIG. 4 shows the diarrhea scores of Experiment Groups 2 and 3 andComparative Group 2.

[0030]FIG. 5 shows the diarrhea scores of Experiment Group 4 andComparative Group 3.

[0031]FIG. 6 shows the diarrhea scores of Experiment Group 5 andComparative Group 4.

[0032]FIG. 7 shows the diarrhea scores of Experiment Groups 6 and 7 andComparative Groups 5 through 7.

[0033]FIG. 8 shows the dry weights of stools from Experiment Group 1aand Comparative Groups 1a and 2a.

[0034]FIG. 9 is a set of photographs showing the biological morphologyof the appearance of mucosas of the small intestines of ExperimentGroups 1a and Comparative Groups 1a and 2a (9-2, 9-1, and 9-3,respectively).

[0035]FIG. 10 is a pair of photographs showing the appearance of stoolsfrom Experiment Group 1b and Comparative Group 1b.

[0036]FIG. 11 is a pair of photographs showing the peripheral appearanceof the anuses of Experiment Group 1b and Comparative Group 1b.

BEST MODE FOR CARRYING OUT THE INVENTION

[0037] As already mentioned, the substance containing proteins andinsoluble dietary fibers as used herein is a component obtained from thegerminated seed of a grass family plant. Grass family plants refer toall the plants that are classified into the grass family. Specifically,they include, but are not limited to, rice, barley, wheat, rye, millet,barnyard grass, corn, and the like. Among them, rice, barley, and ryeare preferred. The germinated seed of a grass family plant may be usedas a raw material in separateing a substance containing proteins andinsoluble dietary fibers with the protein content being 10 to 70% byweight and the insoluble dietary fiber content 20 to 70% by weight, andpreferably with the protein content being 10 to 60% by weight and theinsoluble dietary fiber content 20 to 50% by weight. To determine theprotein content as used herein, then multiplied the nitrogen content isfirst obtained using the Kjeldahl method and by 6.25 which is a proteinconversion coefficient. The content of dietary fibers is the sum of theamounts of all dietary fibers present that are determined based on themethod shown in “Dietary Fiber”, Innami and Kiriyama, eds., pp.38 -40,1989, Daiichi Shuppan.

[0038] Specific methods for producing such the substance containingproteins and insoluble dietary fibers include the following. i.e, thehusks of germinated seeds of a grass family are gradually scraped off(until the weight becomes about 90% that of the original raw material) ,and then the fractions containing the aleurone layer and the germ aregradually scraped off until the residue is endosperm only (until theweight becomes about 80% that of the original raw material), and thealeurone layer-germ fraction is obtained. The weights mentioned aboveare just guide figures and the fractionation is preferably carried outwith the aleurone layer etc. are examined under an electron microscope.

[0039] Economically, it is preferred to use brewer's grains, which arethe barley malt remaining after use as the raw material of beer. As aspecific example of the method for obtaining the substance containingproteins and insoluble dietary fibers from brewer's grains, the onedescribed in Japanese Post-Unexamined Patent Publication (Kokoku) No.4-31666 can be used. Thus, the brewer's grains in the wet state aretreated by pressing and milling and the pressed and milled product thusobtained may be sieved in the presence of water. The fraction that haspassed through the sieve is the substance containing proteins andinsoluble dietary fibers. More particularly, brewer's grains in the wetstate are first treated by pressing and milling. Although any pulverizercan be used that has a structure capable of exerting a compressive forceon the raw material to be treated for pressing and milling of thebrewer's grains, a roll mill is specifically preferred. The gap betweenthe rolls is 0.05 to 2 mm, preferably 0.1 to 0.3 mm. When the brewer'sgrains are to be pressed and milled, a water content in the brewer'sgrains is preferably adjusted to 65% or higher. Then the pressed andmilled product is sieved in the presence of water. In this sieving, thehusk remains on the sieve and the substance containing proteins andinsoluble dietary fibers passes through it. The size of the openings inthe sieve is 5 to 50 mesh, preferably 20 to 50 mesh. In order to obtainefficiently the fraction containing proteins and insoluble dietaryfibers, the above-mentioned pressing and milling and sieving operationsare preferably repeated 2 to 5 times. The fractions thus obtained as theundersizes through the sieve are usually dried before use. The dryingmethods include, but are not limited to, drying with warm air at atemperature of 50 to 100° C., and lyophilization. In some cases, thefractions may be used in the wet state, where their water content ispreferably adjusted to about 10 to 90%.

[0040] The dispersibility of the material into water may be enhanced byconverting the contained proteins into smaller molecules using anenzyme, etc. For example, the substance containing proteins andinsoluble dietary fibers is dispersed in water of pH 9, which is thensubjected to a thermal dispersing treatment such as autoclaving. Afterthe temperature of the dispersion has risen to 50° C., alkali protease(such as Alkalase by Novo) is added to about 0.02% followed byincubation for about 24 hours, where upon the proteins are converted tosmaller molecules with an average peptide chain length of about 5 to 8.The average peptide chain length can be determined by the TNBS method(Nakamura et al., Nippon Shokuhin Kogyo Gakkaishi 38: 377-383, 1991).

[0041] Furthermore, the content of the insoluble dietary fibers may beenhanced by removing some of the protein fraction. In one method, forexample, the substance containing proteins and insoluble dietary fibersis treated by artificial digestion (Matoba et al., Journal of JapaneseSociety of Food and Nutrition, 34: 415-421, 1981). In the method, thefraction of proteins and insoluble dietary fibers is sequentiallyreacted in a pepsin-hydrochloric acid solution and then in apancreatin-hydrochloric acid solution in order to yield an undigestedfraction. By this treatment, the content of dietary fibers can beenhanced to as high as 70% by weight. However, this is not the soleexamples of the methods that can be applied.

[0042] Oral administration of the substance containing proteins andinsoluble dietary fibers can not only alleviate severe diarrhea derivedfrom ulcerative colitis, but daily intake of the substance can increaseresistance to ulcerative colitis itself. For this purpose, the substancecontaining proteins and insoluble dietary fibers is preferally taken inat least 1 g, more preferably at least 12 g, per day. Since thesubstance containing proteins and insoluble dietary fibers is derivedfrom the germinated seed of a grass family plant, it will do no harmeven if it is taken excessively and, hence, it can be adopted in dailyeating programs. The administration or intake of the substance may betimed before, between , or after meals.

[0043] The substance containing proteins and insoluble dietary fibersmay be used to prevent or reduce the side effects of cancer treatments.The methods of cancer treatment include the radiation therapy in which aradiation such as gamma rays are applied, the chemotherapy in whichanti-cancer drugs such as methotrexate and fluorouracil areadministered, and combinations of these methods. The side effects ofcancer treatments include damages to intestinal mucosa and the resultantdiarrhea. In accordance with the present invention, the substancecontaining proteins and insoluble dietary fibers may be given at leastone day, preferably one week, before cancer treatment. Alternatively,the substance containing proteins and insoluble dietary fibers may begiven concomitantly with or after cancer treatment. The daily dosage foradults is 1 g or more, preferably 12 g or more. The substance is derivedfrom the germinated seed of a grass family plant, so it will do no harmeven if it is taken excessively. Accordingly, it is preferablyadministered continuously during the cancer treatment.

[0044] Oral intake of the substance containing proteins and insolubledietary fibers can promote improvement in the bowel movement of apatient who has undergone massive ablation of the bowels or a patientwho wears an artificial anus. For this purpose, the proteins andinsoluble dietary fibers are preferably taken in at least 4 g, morepreferably at least 12 g, per day, Since the proteins and insolubledietary fibers are components derived from the germinated seed of agrass family plant, it will do no harm even if it is taken excessively.

[0045] The substance containing proteins and insoluble dietary fibersaccording to the present invention may be added to foods and beverages.Examples of the foods and beverages to which the substance containingproteins and insoluble dietary fibers according to the present inventioncan be added include those containing natural products or processedproducts as the derivatives. The substance can be added in an amount of0.01 to 1 g per gram of food or beverage. The substance containingproteins and insoluble dietary fibers according to the present inventioncan be added to foods and beverages in various dosage forms such assolutions, suspensions, powders, granules, capsules, and the like.

[0046] The substance containing proteins and insoluble dietary fibersaccording to the present invention which can be formulated as apharmaceutical preparation. In this case, the mode of drugadministration is in no way but exemplary routes of administration maybe peroral, enteral and the like. In the case of oral or enteraladministration, the substance containing proteins and insoluble dietaryfibers can be administered as it is or alternatively it can beadministered in the form of a solution, a suspension, powder, granule, atablet, a capsule, and the like in combination with a pharmaceuticallyacceptable excipient. In this case, the preparation contains the abovesubstance containing proteins and insoluble dietary fibers according tothe present invention in a suitable amount of 1 to 50%, preferably 1 to30%. Examples of the excipient include: sugars such as lactose,sucroseand glucose; starch, inorganic substances such as calcium carbonate andcalcium sulfate; and other commonly used substances such as crystallinecellulose, distilled water, purified water, sesame oil, soybean oil,corn oil, olive oil, and cotton seed oil. In formulating pharmaceuticalpreparations, additives such as binders, lubricants, dispersants,suspending agents, emulsifiers, diluents, buffers, anti-oxidants,anti-bacterial agents can be used. Other pharmaceutical preparations maybe mixed or used in combination. The above preparations may besterilized.

[0047] The present invention is now explained in further detail withreference to the following examples. It should be noted, however, thatthe scope of the present invention is not limited by these examples inany way. Unless otherwise specified, % means present by weight in theexamples.

[Formulation Example 1]

[0048] Brewer's grains in the wet state (water content: 77.6% by weight)were pressed and milled (the rotating speed of the roller: 100 rpm, thegap between rolls: 0.1 mm), and then screened using a 50 mesh sieve. Thefraction that passed through the sieve was dried using a steam dryer andthen crushed to give a substance containing proteins and insolubledietary fibers. The analytical values of the substance were as shown inTable 1 and 2.

[0049] Crude protein as shown in Table 1 was determined using Kjeldahlmethod (the coefficient of nitrogen conversion to protein was set at6.25), crude fat was determined using the soxhlet extraction method withdiethyl ether as the extraction solvent, the ash content was determinedby the direct in cineration method with the sample placed in a crucible,and the dietary fiber content was determined by calculating the sum ofthe amounts of hemicellulose, cellulose, and lignin shown in Table 2.

[0050] The analyses of hemicellulose, cellulose, and lignin shown inTable 2 were carried out based on the method described in “DietaryFiber,” Innami and Kiriyama eds., pp.38 -40, 1989, Daiichi Shuppan.TABLE 1 Table of Ingredients (% by weight) Crude protein 53.4 Crude fat12.6 Ash content 2.0 Dietary fiber 32.1

[0051] TABLE 2 Composition of Dietary fiber (%) Hemicellulose 50.0Cellulose 24.0 Lignin 26.0

[Formulation Example 2]

[0052] With the gap between rolls adjusted to 2 mm, a different kind ofbrewer's grains was treated as in Formulation Example 1 to give thesubstance containing proteins and insoluble dietary fibers which wasderived from the brewer's grains and which contained different amountsof proteins and insoluble dietary fibers. The analytical values of thesubstance are shown in Table 3. TABLE 3 Composition of the substancecontaining proteins and insoluble dietary fibers as obtained inFormulation Example 3 Ingredients (% by weight) Crude protein 56.1 Crudefat 12.0 Ash content 2.8 Dietary fiber 28.9

[Formulation Examples 3 and 4]

[0053] Using the same method as shown in Formulation Example 2 exceptthat a different kind of brewer's grains was used as the raw materialand that the conditions of the roll mill and screening in water weremodified, the substance containing proteins and insoluble dietary fiberswas obtained which was derived from the brewer's grains and whichcontained different amounts of proteins and insoluble dietary fibers.The analytical values of the substance are shown in Table 4. TABLE 4Composition of proteins and insoluble dietary fibers of FormulationExamples 3 and 4 (% by weight) Ingredients Formulation Example 3Formulation Example 4 Crude protein 34.9 60.0 Crude fat 12.0 12.0 Ashcontent  3.0  2.8 Dietary fiber 50.1 25.0

[Formulation Example 5]

[0054] Starting with the substance containing proteins and insolubledietary fibers which obtained in Formulation Example 1, a materialhaving good dispersibility in water was prepared.

[0055] Fifty grams (in the dry state) of the substance containingproteins and insoluble dietary fibers as obtained in Formulation Example1 was weighed, adjusted to pH 9 with sodium hydroxide and furtheradjusted to make 1 liter. The mixture was subjected to a heat treatmentsuch as autoclaving (121° C., 10 min) and then cooled to 50° C.Thereafter, about 0.02% of alkaline protease (such as Alkalase by Novo)followed by incubation for 24 hours. Subsequently, thermal-denaturing ofthe enzyme and lyophilization were conducted. The lyophilized productwas crushed to yield a substance having good dispersibility in water.The average peptide chain length of the substance was determined by theTNBS method (Nakamura et al., Nippon Shokuhin Kogyo Gakkaishi 38:377-383, 1991) and found to be about 5 to 8.

[Formulation Example 6]

[0056] A material with an enhanced content of dietary fibers wasprepared from the substance containing proteins and insoluble dietaryfibers that had been obtained in Formulation Example 1.

[0057] One hundred grams of the fraction obtained in Formulation Example1 was taken as a sample; 1600 ml of an enzyme solution (0.087%pepcin/0.096 N HCl+0.03 M NaCl) and, furthermore, 1200 ml of 0.1 MNaHCO3 were added. The mixture was thermally denatured by incubating at37° C. for 4 hours. Washing with water was performed until a neutralfiltrate came out; thereafter, 1000 ml of an enzyme solution (2.17%pancreatin/Tris-HCl 0.5M, pH 8) and, furthermore, 4000 ml of 0.01 N HClwere added. The mixture was thermally denatured by incubating at 37° C.for 24 hours or longer (26 hours in this case); it was then washed,filtered, lyophilized and crushed. By these treatments, the proteincontent became 15% and the dietary fiber content 70%.

[Test Example 1]

[0058] The substance containing proteins and insoluble dietary fibersthat was prepared in Formulation example 1 was tested to determinewhether it could treat the severe diarrhea from ulcerative colitis orabnormalities in the mucosa of colon.

Materials and Methods

[0059] As the test animal, male SD rats (3-week old, about 50 g) thathad been acclimated to the experimental condition by a preparatorybreeding with the solid feed (CE-2, Nippon clea) for one week were usedin groups of five animals each. The feeds used in Test Example 1 were asshown in Table 5.

[0060] Ulcerative colitis was experimentally developed by giving theanimal the feed mixed with dextran sodium sulfate. This is amodification of Iwanaga's method (Journal of Gastroenterology 29:430-438, 1994). The substance containing proteins and insoluble dietaryfibers was also given ad libitum in a mixture with the feed. Afterbreeding for 5 days, the appearances of the stool and the anus wereexamined and the state of diarrhea was evaluated by scoring. Then thecolon was excised and fixed in 10% formalin buffer to prepare sectionsof the mucosa, and the microscopic view of the mucosa were alsoexamined.

[0061] Table 5 Composition of the feed

[0062] The basic feed was prepared so as to contain 10% of protein fromcasein, 5% of lipid from corn oil, 1% of vitamin, 3.5% of minerals, and0.2% of choline chloride. The weight was adjusted using corn starch.

Comparative Group 1

[0063] The basic feed was supplemented with casein and cellulose toprovide the protein and dietary fiber contents that would be given byadding 10% of the substance containing proteins and insoluble dietaryfibers according to Formulation Example 1. Then, dextran sodium sulfatewas added at 4% to induce ulcerative colitis. Experiment Group 1: Afterthe substance containing proteins and insoluble dietary fibers accordingto Formulation Example 1 was added to the basic feed at 10%, dextransulfate sodium was added at 4% to induce ulcerative colitis. ComparativeExperiment Group 1 Group 1 (% by weight) Casein 14.6 10 Mineral mix*¹3.5 3.5 Vitamin mix*² 1 1 Starch 68.7 66.3 Corn oil 5 5 Cellulose 3 Thesubstance containing proteins 10 and insoluble dietary fibers accordingto Formulation example 1 Dextran sodium sulfate 4 4 Choline chloride 0.20.2

Results

[0064] The results are shown in FIG. 1 through 3. FIG. 1 shows thediarrhea scores of Experiment Group 1 and Comparative Group 1. Thediarrhea score of FIG. 1 is the average of scores assigned to the ratsin each group on the basis of the following 6-level rating: 0, normal;1, slightly soft; 2, considerably soft; 3, stools cannot be picked up;4, diarrhea+slightly eroded anus; 5, diarrhea+badly eroded anus. FIG. 2shows the appearance of stools from Comparative Group 1 (2-1) andExperiment Group 1 (2-1). FIG. 3 shows the appearance of the colonmucosas of Experiment Group 1 and Comparative Group 1. As shown in FIG.1 through 3, amelioration in diarrhea and prevention of damages in thecolon mucosa were observed only when the substance containing proteinsand insoluble dietary fibers was taken in. Similarly, diffusion of anuswas also prevented only when the substance containing proteins andinsoluble dietary fibers was taken in. In both of FIG. 2 and FIG. 3,only one case is illustrated for each experiment group, but normalimages of the mucosa were observed in 4 out of 5 animals in the groupthat took the substance containing proteins and insoluble dietaryfibers. As can be seen from this example, the substance containingproteins and insoluble dietary fibers effectively promotes ameliorationof symptoms.

[0065] Similar results have been obtained for the substance containingproteins and insoluble dietary fibers according to the FormulationExamples 2 through 4.

[Test example 2]

[0066] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 1 was tested to determine whether itcould treat severe diarrhea associated with ulcerative colitis orabnormalities in the mucosa of colon.

Materials and Methods

[0067] These were basically the same as in Test Example 1, butulcerative colitis was developed by 5-day breeding of all rats with thefeed of Comparative Group 1 (the control group) described in TestExample 1; then, the rats were bred for 3 days, with the feed containingno dextran sodium sulfate as shown in Table 6 and the degree of recoveryfrom diarrhea was evaluated by scoring in a similar manner to TestExample 1. From the results of Test Example 1, it was concluded that inthe cases of complete recovery from diarrhea, the mucosa of the colonwas very close to the normal state.

[0068] Table 6 Composition of the feed

[0069] The method of preparing the basic feed was the same as in TestExample 1.

[0070] Comparative Group 2: The basic feed was supplemented with caseinand cellulose to provide the protein and dietary fiber contents thatwould be given by adding 10% of the substance containing proteins andinsoluble dietary fibers according to Formulation Example 1.

[0071] Experiment Group 2: The substance containing proteins andinsoluble dietary fibers according Formulation Example 1 was added tothe basic feed at 10%.

[0072] Experiment Group 3: Salazosulfapyridine (salazopyrine:manufactured by Green Cross K.K.) was added to the feed of ComparativeGroup 2 at 0.5 g/kg feed (so that the approximate dose would be equal to50 mg/kg body weight). The dose of salazosulfapyridine was decided byreferring to Folia. Pharmacol. Jpn. 102: 343-350 (1993). Com- Ex-parative periment Experiment Group 2 Group 2 Group 3 (% by weight)Casein 14.6 10 14.6 Mineral mix*¹ 3.5 3.5 3.5 Vitamin mix*² 1 1 1 Starch72.7 70.3 72.65 Corn oil 5 5 5 Cellulose 3 3 The substance containingproteins 10 and insoluble dietary fibers according to FormulationExample 1 Salazopyrine 0.05 Choline chloride 0.2 0.2 0.2

Results

[0073] The results are shown in FIG. 4. FIG. 4 shows the diarrhea scoresof Experiment Groups 2 and 3, and Comparative Group 2. Evaluation byscoring was made in a similar manner to Test Example 1. As shown in FIG.4, recovery from diarrhea was very fast in the group that was given thesubstance containing proteins and insoluble dietary fibers. The degreewas significantly better than from salazosulfapyridine, a drug currentlyused as a therapeutic drug for ulcerative colitis. The presentexperiment has shown that the administration of the substance containingproteins and insoluble dietary fibers can effectively promoteamelioration of symptoms even if it is taken after development ofulcerative colitis. Similar results are confirmed for the substancecontaining proteins and insoluble dietary fibers according toFormulation Examples 2 through 4.

[Test Example 3]

[0074] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 5 was tested to determine whether itcould treat severe diarrhea associated with ulcerative colitis orabnormalities in the mucosa of colon.

Materials and Methods

[0075] These were basically the same as in Test Example 1. The feedsused in Test example 3 were as shown in Table 7. After breeding for 5days, the appearances of the stool and the anus were examined and theappearance of diarrhea was evaluated by scoring.

[0076] Table 7 Table of feed composition

[0077] The method of preparing the basic feed was the same as in TestExample 1.

[0078] Test Example Group 3: The basic feed was supplemented with caseinand cellulose to provide the protein and dietary fiber contents thatwould be given by adding 10% of the substance containing proteins andinsoluble dietary fibers according to Formulation Example 5. Then,dextran sodium sulfate was added thereto at 4% to induce ulcerativecolitis.

[0079] Experiment Group 4: The substance containing proteins andinsoluble dietary fibers of Formulation Example 5 was added to the basicfeed at 10%, and dextran sulfate sodium was added at 4% to induceulcerative colitis. Comparative Experiment Group 3 Group 4 (% by weight)Casein 14.6 10 Mineral mix*¹ 3.5 3.5 Vitamin mix*² 1 1 Starch 68.7 66.3Corn oil 5 5 Cellulose 3 The substance containing proteins 10 andinsoluble dietary fibers according to Formulation example 5 Dextransodium sulfate 4 4 Choline chloride 0.2 0.2

Results

[0080] The results are shown in FIG. 5. FIG. 5 shows the diarrhea scoresof Experiment Group 4, and Comparative Group 3. Evaluation by scoringwas made in a similar manner to Test example 1. As shown in FIG. 5, thesubstance containing proteins and insoluble dietary fibers ofFormulation Example 5 has also shown a very marked amelioration of mainsymptoms of ulcerative colitis.

[Test Example 4]

[0081] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 6 was tested to determine whether itcould treat severe diarrhea associated with ulcerative colitis orabnormalities in the mucosa of colon.

Materials and Methods

[0082] As the test animal, male Std-ddY mice (6-week old, about 30 g)that had been acclimated to the experimental condition by a preparatorybreeding with the solid feed (CE-2, Nippon clea) for one week were usedin groups of five animals each. The feeds used in Test Example 4 were asshown in Table 8.

[0083] Ulcerative colitis was experimentally developed by giving theanimal the feed mixed with dextran sodium sulfate. This is amodification of Iwanaga's method (Journal of Gastroenterology 29:430-438, 1994). The substance containing proteins and insoluble dietaryfibers was also given ad libitum in a mixture with the feed. Afterbreeding for 3 days, the appearances of the stool and the anus wereexamined on day 4 and the state of diarrhea was evaluated by scoring.

[0084] Table 8 Composition of the feed

[0085] The method of preparing the basic feed was the same as in TestExample 1.

[0086] Comparative Group 4: Dextran sodium sulfate was added to thebasic feed at 4% to induce ulcerative colitis.

[0087] Experiment Group 5: After the substance containing proteins andinsoluble dietary fibers according to Formulation Example 6 was added tothe basic feed at 5%, dextran sodium sulfate was added at 4% to induceulcerative colitis. Comparative Experiment Group 1 Group 1 (by weight)Casein 10 10 Mineral mix*¹ 3.5 3.5 Vitamin mix*² 1 1 Starch 73.3 71.3Corn oil 5 5 Cellulose 3 4 The substance containing proteins 5 andinsoluble dietary fibers according to Formulation Example 6 Dextransodium sulfate 4 4 Choline chloride 0.2 0.2

Results

[0088] The results are shown in FIG. 6. FIG. 6 shows the diarrhea scoresof Experiment Group 5 and Comparative Group 4. Evaluation by scoring wasmade in a similar manner to Test Example 1. As shown in FIG. 6, thesubstance containing proteins and insoluble dietary fibers according toFormulation Example 6 has also shown a very effective amelioration ofmain symptoms of ulcerative colitis.

[Test Example 5]

[0089] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 1 was tested to determine whether itcould prevent or ameliorate severe damages on the mucosa of colon anddiarrhea when anti-cancer agents were being used.

Materials and Methods

[0090] Male SD rats (4-week old), after preparatory breeding with thesolid feed (CE-2, Nippon clea) for one week, were divided into groups offive animals each (body weight at the time of grouping was 136.2 g).They were then bred for one week by giving ad libitum the feed shown inTable 9. On day 7, 8 and 9, methotrexate (manufactured by NipponLederle) was given intraperitoneally at 5 mg/kg body weight. The animalswere further bred for 5 days, and on day 5, after fasting for 4 hours,the jejunum (portions at 15 to 30 cm from the pylorus) was taken outunder anesthesia with urethane. It was then fixed in formalin andembedded with paraffin and stained with hematoxylin-eosin formicroscopic observation.

[0091] The following comparative examples were set up: 1) a comparativecontrol group (cellulose was used as a dietary fiber) and 2) a group towhich glutamin commonly held as an ideal nutrient for intestinal mucosawas added. On the last three days, stools were collected and their dryweight was measured.

[0092] Table 9 Table of feed composition

[0093] The method of preparing the basic feed was the same as in

Test Example 1.

[0094] Comparative Group 1a: The basic feed was supplemented with caseinand cellulose to provide the protein and dietary fiber contents thatwould be given by adding 10% of the substance containing proteins andinsoluble dietary fibers according to Formulation Example 1.

[0095] Experiment Group 1a: The substance containing proteins andinsoluble dietary fibers according to Formulation Example 1 was added tothe basic feed at 10%.

[0096] Experiment Group 2a: Glutamin was added to the basic feed at1.13%.

[0097] This is equivalent to the content of glutamin obtained when thesubstance containing proteins and insoluble dietary fibers was added tothe feed at 10%. Furthermore, casein and cellulose were added so thatthe weights of proteins and dietary fibers would be the same as inExperiment Group 1a. Com- Ex- Ex- parative periment periment Group 1aGroup 1a Group 2a (% by weight) Casein 14.6 10 13.47 Mineral mix*¹ 3.53.5 3.5 Vitamin mix*² 1 1 1 Starch 72.7 70.3 72.7 Corn oil 5 5 5Cellulose 3 3 The substance containing proteins 10 and insoluble dietaryfibers according to Formulation Example 1 Glutamin 1.13 Choline chloride0.2 0.2 0.2

Results

[0098] The results are shown in FIGS. 8 and 9. FIG. 8 shows the dryweight of the stools from Experiment Group 1a and Comparative Group 1aand 2a. Comparative Groups 1a and 2a had severe diarrhea. FIG. 9 showsthe appearance of the mucosas of the small intestines of ExperimentGroup 1a and Comparative Groups 1a and 2a (2-2, 2-1, and 2-3,respectively). As shown in FIGS. 8 and 9, excretion of the stool wasnormal in Experiment Group 1a that took the substance containingproteins and insoluble dietary fibers, but a reduction in the dry weightof stool excretion was observed in Comparative Groups 1a and 2a due todiarrhea. In the examination of the mucosa of the intestine, images ofnormal mucosa were observed only when the substance containing proteinsand insoluble dietary fibers was taken and this suggests that thesubstance protects the intestinal mucosa from side effects of theanti-cancer agent.

[Test example 6]

[0099] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 1 was tested to determine whether itcould reduce or prevent the damage that would otherwise be caused on theintestinal mucosa by radiation exposure.

Materials and Methods

[0100] Male ST-Wistar rats (3-week old), after preparatory breeding withthe casein-sucrose diet (the basic feed or Comparative Group 3a as shownin Table 10) for one week, were divided into groups of seven animalseach. Subsequently, the animals were fed the test feed shown in Table 10for 10 days, and on day 11 they were exposed to a radiation of 10 Gy(from 60 Co) in a localized area (lower abdomen). They were bred for 8more days and the effect of the substance containing proteins andinsoluble dietary fibers to prevent the damage on the intestinal mucosadue to radiation exposure was evaluated in terms of the survival rate.

[0101] Table 10 Table of feed composition

[0102] Comparative Group 3a:The sucrose basic feed group with 20% caseinand 0% dietary fiber

[0103] Experiment Group 2a: The substance containing proteins andinsoluble dietary fibers prepared in Formulation example 1 was added tothe sucrose basic feed at 10% and its sucrose content was accordinglyreduced. Comparative Experiment Group 3a Group 2a (% by weight) Casein20 20 Mineral mix 3.5 3.5 Vitamin mix 1 1 Sucrose 70.3 60.3 Corn oil 5 5The substance containing proteins 10 and insoluble dietary fibersaccording to Formulation example 1 Choline chloride 0.2 0.2

Results

[0104] As shown in Table 11, a reduction in mortality was observed onlywhen the substance containing proteins and insoluble dietary fibers wastaken by the test animals. This is believed to result from the reductionin the radiation damage on the intestinal mucosa. TABLE 11 Survival rateon day 8 of radiation exposure Comparative Experiment Group 3a Group 2a(No. of viable animals/total No. tested) (5/7) (7/7)

[Test example 7]

[0105] The substance containing proteins and insoluble dietary fibersprepared in Formulation example 1 was tested to determine whether itwould permit normal stool formation when a massive ablation of the lowergastrointestinal tract (the large intestine) was carried out.

Materials and Methods

[0106] As the test animal, male SD rats (4-week old, about 50 g) thathad been acclimated to the experimental condition by a preparatorybreeding with the feed shown in Table 12 for one week were used ingroups of 10 animals each. The large intestine was removed in such a waythat the anus and the large intestine within the pelvis were leftintact; then, an end-to-end anastomosis of the small intestine wasconducted. After a recovery period of 2 days, the animals were givenagain the feed shown in Table 12 and bred for one week, and theappearance of the stool and the anus were examined.

[0107] Table 12 Table of feed composition

[0108] The method of preparing the basic feed was the same as in TestExample 1.

[0109] Comparative Group 1b: The basic feed was supplemented with caseinand cellulose to provide the protein and dietary fiber contents thatwould be given by adding 10% of the substance containing proteins andinsoluble dietary fibers according to Formulation Example 1.

[0110] Experiment Group 1b: The substance containing proteins andinsoluble dietary fibers according to Formulation Example 1 was added tothe basic feed at 10%. Comparative Experiment Group 1b Group 1b (% ofweight) Casein 14.6 10 Mineral mix*¹ 3.5 3.5 Vitamin mix*² 1 1 Starch72.7 70.3 Corn oil 5 5 Cellulose 3 The substance containing proteins 10and insoluble dietary fibers according to Formulation Example 1 Cholinechloride 0.2 0.2

Results

[0111] Examination of the stool and the anus was conducted on the lastday of breeding and the results are shown in FIGS. 10 and 11: fairlysevere diarrhea with liquid stools was noted in Comparative Group 1bwhereas the stool had an almost normal appearance in Experiment Group1b.

[0112] Examination of the periphery of the anus revealed that severeerosion due to diarrhea was noted in Comparative Group 1b whereas noerosion was observed to give an almost normal appearance in ExperimentGroup 1b because of the component of interest in Formulation Example 1.

[0113] The substance containing proteins and insoluble dietary fiberspermitted normal stool formation very effectively after ablation of thegastrointestinal tract.

[Formulation Example 7]

[0114] After accurate weight measurement, dry barley malt was scrapedfrom the husk surface with TDB2A, a rice whiting machine for brewery use(manufactured by Satake Seisakusho; working rotational speed, 500 rpm) .The scraped malt grains which weighed 88 - 80% of the initial barleymalt were obtained as the substance containing proteins and insolubledietary fibers. Its protein and dietary fiber contents were 11.5% and30%, respectively.

[0115] For comparison, ungerminated barley was fractionated under thesame conditions as described above. The resulting substance containingproteins and insoluble dietary fibers had protein and dietary fibercontents of 16.3% and 31.5%, respectively.

[Formulation Example 8]

[0116] Germinated rice was dried and dehusked. After accurate weightmeasurement, the dehusked grains were scraped from the surface withTDB2A, a rice whiting machine for brewery use (manufactured by SatakeSeisakusho; working rotational speed, 500 rpm). The scraped rice grainswhich weighed 100 - 95% of the initial rice grains were obtained as thesubstance containing proteins and insoluble dietary fibers. Its proteinand dietary fiber contents were 17.6% and 21.6%, respectively.

[0117] For comparison, ungerminated brown (dehusked) rice wasfractionated under the same conditions as described above. The resultingsubstance containing proteins and insoluble dietary fibers had proteinand dietary fiber contents of 16.3% and 28.5%, respectively.

[Test Example 8]

[0118] The effect of the substance containing proteins and insolubledietary fibers prepared in Test Examples 7 and 8 was investigated.

Materials and Methods

[0119] The same as in Test Example 1. The feeds used in Test Example 8were as shown in Table 13.

[0120] Table 13 Table of feed composition

[0121] The method of preparing the basic feed was the same as in TestExample 1.

[0122] Comparative Group 5: Same as the feed for Experiment Group 6,except that no substance containing proteins and insoluble dietaryfibers was added and that it was supplemented with casein and celluloseto provide the corresponding protein and dietary fiber contents. Itcontained 3.5% of dextran sodium sulfate in order to induce ulcerativecolitis.

[0123] Comparative Group 6: The substance containing proteins andinsoluble dietary fibers which was derived from “ungerminated barley”and prepared in Formulation Example 7 was added to the basic feed at10%, and dextran sodium sulfate was added at 3.5% in order to induceulcerative colitis.

[0124] Experiment Group 6: The substance containing proteins andinsoluble dietary fibers which was derived from barley malt and preparedin Formulation Example 7 was added to the basic feed at 10%, and dextransodium sulfate was added at 3.5% in order to induce ulcerative colitis.

[0125] Comparative Group 7: The substance containing proteins andinsoluble dietary fibers which was derived from “ungerminated brownrice” and prepared in Formulation Example 8 was added to the basic feedat 10%, and dextran sodium sulfate was added at 3.5% in order to induceulcerative colitis.

[0126] Experiment Group 7: The substance containing proteins andinsoluble dietary fibers which was derived from germinated rice grainsand prepared in Formulation Example 8 was added to the basic feed at10%, and dextran sodium sulfate was added at 3.5% in order to induceulcerative colitis. Comp. Comp. Exp. Comp. Exp. Group 5 Group 6 Group 6Group 7 Group 7 (% by weight) Casein 14.6 13.0 13.3 13.0 12.8 Mineralmix 1 1 1 1 1 Vitamin mix 3.5 3.5 3.5 3.5 3.5 Starch 69.4 64.0 63.5563.85 63.36 Cellulose 3 0.15 0.15 0.84 Corn oil 5 5 5 5 5 The substancecontaining proteins 10 and insoluble dietary fibers derived fromungerminated barley according to Formulation Example 7 The substancecontaining proteins 10 and insoluble dietary fibers derived from barleyaccording to Formulation Example 7 The substance containing proteins 10and insoluble dietary fibers derived from ungerminated brown riceaccording to Formulation Example 8 The substance containing proteins 10and insoluble dietary fibers derived from germinated rice according toFormulation Example 8 Dextran sodium sulfate 3.5 3.5 3.5 3.5 3.5

Results

[0127] The results of the investigation on Experiment Groups 6 and 7,and Comparative Groups 5 through 7 described in Table 13 are shown inFIG. 7. The diarrhea scores in FIG. 7 were evaluated in the same manneras in Test Example 1.

[0128] As shown in FIG. 7, the diarrhea score decreased only when thesubstances containing proteins and insoluble dietary fibers that derivedfrom the germinated barley and the germinated rice was taken by theanimals. Therefore, it was demonstrated that both of these substancescontaining proteins and insoluble dietary fibers can prevent the severediarrhea associated with ulcerative colitis.

[0129] In Test Example 8, none of the experiment groups showedmeasurable changes in body weight.

[Formulation Example 9]

[0130] After accurate weight measurement, dried and dehusked rye maltwas scraped from the grain surface with TDB2A, a rice whiting machinefor brewery (manufactured by Satake Seisakusho; working rotationalspeed, 500 rpm).

[0131] The scraped rye malt grains which weighed 100 - 96% of theinitial rye malt grains were obtained as the substance containingproteins and insoluble dietary fibers. Its protein and dietary fibercontents were 16.3% and 35%, respectively.

[Formulation Example 10]

[0132] After accurate weigh measurement, dried and dehusked rye wasscraped from the grain surface with TDB2A, a rice whiting machine forbrewery use (manufactured by Satake Seisakusho; working rotationalspeed, 500 rpm). The scraped rye grains which weighed 100 - 96% of theinitial rye grains were obtained as the substance containing proteinsand insoluble dietary fibers. Its protein and dietary fiber contentswere 12.9% and 35%, respectively.

[0133] The analytical values of these substances are as shown in Table14. TABLE 14 Table of ingredients Formulation Example 9 FormulationExample 10 (% by weight) Crude protein 16.3 12.9 Crude fat 3.8 4.2 Ashcontent 2.6 3.7 Dietary fiber 35.0 35.0

[Test Example 9]

[0134] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 9 was tested to determine whether itcould treat the severe diarrhea associated with ulcerative colitis orabnormalities in the mucosa of colon. The same test was conducted withthe fractions obtained in Formulation Example 10.

Materials and Methods

[0135] As the test animal, male SD rats (3-week old, about 50 g) thathad been acclimated to the experimental condition by a preparatorybreeding with a solid feed for one week were used in groups of fiveanimals each. The feeds used in Test Example 9 were as shown in Table15.

[0136] The substances containing proteins and insoluble dietary fibersprepared in Formulation Examples 9 and 10 were also given ad libitum inadmixture with the feed. After breeding for 5 days, the appearance ofthe stool and the anus was examined and the severity of diarrhea wasevaluated by scoring. Then the colon was ablated totally and fixed in10% formalin to prepare sections of the mucosa, and the images of themucosa were also examined. The results were, as shown in Table 16; theamelioration of diarrhea and the prevention of erosion in the anus anddamage on the mucosa of the large intestine were possible only when thesubstance containing proteins and insoluble dietary fibers prepared inFormulation Example 9 was taken by the animals.

[0137] The severity of diarrhea was classified into 6 levels: 0, normal;1, slightly soft; 2, considerably soft; 3, stools could not be pickedup; 4, diarrhea+slightly eroded anus; 5, diarrhea+badly eroded anus.Also, the damage on the mucosa of the large intestine was scored inaccordance with the method of Morris, in which the larger number means aseverer damage.

[0138] Table 15 Table of feed composition

[0139] The method of preparing the basic feed was the same as in TestExample 1.

[0140] Comparative Group 1c: The basic feed was supplemented with caseinand cellulose to provide the protein and dietary fiber contents thatwould be given by adding 10% of the substance containing proteins andinsoluble dietary fibers according to Formulation Example 1. Dextransodium sulfate was added at 4% to induce ulcerative colitis.

[0141] Comparative Group 2c: The substance containing proteins andinsoluble dietary fibers according to Formulation Example 10 was addedto the basic feed at 10%, and dextran sodium sulfate was added at 4% inorder to induce ulcerative colitis. Casein and cellulose were also addedto give the same protein and dietary fiber contents as in ComparativeGroup 1c.

[0142] Experiment Group 1c: The substance containing proteins andinsoluble dietary fibers according to Formulation Example 9 was added tothe basic feed at 10%, and dextran sodium sulfate was added at 4% inorder to induce ulcerative colitis. Casein and cellulose were also addedto give the same protein and dietary fiber contents as in ComparativeGroup 1c. Comparative Experiment Experiment Group 1c Group 2c Group 1c(% by weight) Casein 14.6 13.3 12.9 Mineral mix*¹ 3.5 3.5 3.5 Vitaminmix*² 1 1.0 1.0 Starch 68.7 63.0 63.4 Corn oil 5.0 5.0 5.0 Cellulose 3.0The substance containing 10 proteins and insoluble dietary fibersaccording to Formula- tion Example 10 The substance containing 10.0proteins and insoluble dietary fibers according to Formula- tion Example9 Dextran sodium sulfate 4.0 4.0 4.0 Choline chloride 0.2 0.2 0.2

Results

[0143] The results of the above experiment were as shown in Table 16.The substance containing proteins and insoluble dietary fibers preparedin Test Example 9 was capable of treating the diarrhea peculiar toulcerative colitis and it was also very effective in preventing thedamage that would otherwise be caused on the mucosa of the largeintestine. TABLE 16 Score of damage on the mucosa Diarrhea score of thelarge intestine Comparative Group 1c 5.0 4.2 Comparative Group 2c 3.85.7 Experiment Group 1c 0.4 2.5

[Formulation Example 11]

[0144] The method shown in Formulation Example 2 was repeated, exceptthat a different kind of brewer's grains was used as the raw materialand that the drying with dry heat (steam dryer) was replaced bylyophilization. Thus, the brewer's grains in the wet state were pressedand milled and then screened using a 50-mesh sieve within water. Thefraction that passed through the sieve was subjected to preliminaryfreezing at −20° C. and then lyophilized according to the standardmethod.

[0145] The results of analysis are shown in Tables 17 and 18. TABLE 17Table of Ingredients (% by weight) Crude protein 50.2 Crude fat 13.7 Ashcontent 1.5 Dietary fiber 33.6

[0146] TABLE 18 Composition of Dietary Fibers % Hemicellulose 69.9Cellulose 23.3 Lignin 6.8

[0147] A test was conducted with the substance as shown below.

Test Example 10

[0148] The substance containing proteins and insoluble dietary fibersprepared in Formulation Example 11 was tested to determine whether itcould treat the severe diarrhea associated with ulcerative colitis orabnormalities in the mucosa of the large intestine.

Materials and Methods

[0149] The test animals and the method of developing ulcerative colitiswere as shown in Test Example 1, except that the feeds used in TestExample 10 were as shown in Table 19 below. TABLE 19 ComparativeExperiment Example 1 Example 1 (% by weight) Casein 14.6 10 Mineralmix*¹ 35 35 Vitamin mix*² 1.0 1.0 Starch 69.2 66.8 Corn oil 5.0Cellulose 3.0 The substance containing proteins 10.0 and insolubledietary fibers according to Formulation Example 11 Choline chloride 0.20.2 dextran sodium sulfate 3.5 3.5

Results

[0150] The results are shown in Table 20. In Experiment Example 1 thediarrhea score decreased and no erosion was observed in the periphery ofthe anus. TABLE 20 Comparative Example 1 5.0 Experiment Example 1 0.2

[0151] It will be readily recognized by a person skilled in the art thataside from the substances tested in the above Test Examples, those whichare derived from the germinated seeds of plants of a grass family whichare within the scope of the present invention would have similareffects.

Industrial Applicability

[0152] The substance derived from the germinated seed of a grass familyplant of the present invention is capable of treating the diarrheaassociated with ulcerative colitis and it is also effective inpreventing disorders of the mucosa of the large intestine. In addition,the substance derived from the germinated seed of a grass family plantof the present invention can effectively prevent damages on theintestinal mucosa and control the ensuing onset of diarrhea, bothsymptoms being severe side effects resulting from administration ofanti-cancer agents. Furthermore, the substance derived from thegerminated seed of a grass family plant of the present invention caneffectively improve the bowel movement of a patient who has undergoneintestinal ablation or a patient with an artificial anus. Moreover, thesubstance derived from the germinated seed of a grass family plant ofthe present invention has no side effects.

What is claimed is:
 1. A substance isolated from the germinated seed ofa grass family plant, said substance containing proteins and insolubledietary fibers.
 2. The substance as claimed in claim 1 wherein the grassfamily plant is selected from the group consisting of rice, barley andrye.
 3. The substance as claimed in claim 1 comprising the fractionscontaining the aleurone layer and the germ of the germinated seed of agrass family plant.
 4. The substance as claimed in claim 1 wherein theproteins have been converted to smaller molecules with an averagepeptide length of about 5 to 8 by enzyme treatment.
 5. The substance asclaimed in claim 1 wherein the content of insoluble dietary fibers hasbeen enhanced by enzyme treatment and filtration.
 6. The substance asclaimed in claim 1 which contains 10 to 70% by weight of proteins and 20to 70% by weight of insoluble dietary fibers.
 7. A pharmaceuticalcomposition comprising as an active ingredient a substance which wasisolated from the germinated seed of a grass family plant and whichcontains proteins and insoluble dietary fibers.
 8. The pharmaceuticalcomposition as claimed in claim 7 wherein the substance which wasisolated from the germinated seed of a grass family plant and whichcontains proteins and insoluble dietary fibers is a sieve passingfraction obtained by sieving the pressed and milled product of brewer'sgrains.
 9. The pharmaceutical composition as claimed in claim 7 for thetreatment of ulcerative colitis.
 10. The pharmaceutical composition asclaimed in claim 7 for preventing or reducing a side effects of a cancertreatment.
 11. The pharmaceutical composition as claimed in claim 10wherein said side effect of a cancer treatment is a damage on theintestinal mucosa.
 12. The pharmaceutical composition as claimed inclaim 10 wherein said side effect of a cancer treatment is diarrhea. 13.The pharmaceutical composition as claimed in claim 10 wherein saidcancer treatment is by irradiation with gamma rays.
 14. Thepharmaceutical composition as claimed in claim 10 wherein said cancertreatment is by administration of an anti-cancer agent.
 15. Thepharmaceutical composition as claimed in claim 10 wherein said cancertreatment is by the combination of administration of an anti-canceragent and irradiation with gamma rays.
 16. A food composition comprisinga substance which was isolated from the germinated seed of a grassfamily plant and which contains proteins and insoluble dietary fibers.17. The food composition as claimed in claim 16 wherein the substancewhich was isolated from the germinated seed of a grass family plant andwhich contains proteins and insoluble dietary fibers is a sieve passingfraction obtained by sieving the pressed and milled product of brewer'sgrains.
 18. The food composition as claimed in claim 16 for a patientwho has undergone intestinal ablation.
 19. The food composition asclaimed in claim 16 for a patient with an artificial anus.
 20. A methodof treating ulcerative colitis, said method comprising administering toa patient with ulcerative colitis an effective amount of a substancewhich was isolated from the germinated seed of a grass family plant andwhich contains proteins and insoluble dietary fibers.
 21. A method ofavoiding or reducing side effects of cancer treatments, said methodcomprising administering to a patient with a cancer an effective amountof a substance which was isolated from the germinated seed of a grassfamily plant and which contains proteins and insoluble dietary fibers.22. A method of improving the bowel movement of a patient who hasundergone intestinal ablation, said method comprising allowing thepatient to take an effective amount of a substance which was isolatedfrom the germinated seed of a grass family plant and which containsproteins and insoluble dietary fibers.
 23. A method of improving thebowel movement of a patient with an artificial anus, said methodcomprising allowing the patient to take an effective amount of asubstance which was isolated from the germinated seed of a grass familyplant and which contains proteins and insoluble dietary fibers.
 24. Useas a pharmaceutical agent of a substance which was isolated from thegerminated seed of a grass family plant and which contains proteins andinsoluble dietary fibers.
 25. Use, for the treatment of ulcerativecolitis, of a substance which was isolated from the germinated seed of agrass family plant and which contains proteins and insoluble dietaryfibers.
 26. Use, for preventing or reducing side effects of cancertreatments, of a substance which was isolated from the germinated seedof a grass family plant and which contains proteins and insolubledietary fibers.
 27. Use as food of a substance which was isolated fromthe germinated seed of a grass family plant and which contains proteinsand insoluble dietary fibers.
 28. Use, for improving the bowel movementof a patient who has undergone intestinal ablation, of a substance whichwas isolated from the germinated seed of a grass family plant and whichcontains proteins and insoluble dietary fibers.
 29. Use, for improvingthe bowel movement of a patient with an artificial anus, of a substancewhich was isolated from the germinated seed of a grass family plant andwhich contains proteins and insoluble dietary fibers.